Compositions and methods for protein production

Abstract

Provided herein are methods for generating a protein product, comprising: providing a plant seed; contacting said plant seed with a protein expression vector that encodes for a protein; transiently expressing said protein in said plant seed using said protein expression vector, and isolating said protein from said plant seed to obtain a protein product.

What is claimed is: 
     
         1 . A method for generating a protein product, the method comprising
 (a) providing a plant seed;   (b) contacting said plant seed with a protein expression vector that encodes for a protein;   (c) transiently expressing said protein in said plant seed using said protein expression vector; and   (d) isolating said protein from said plant seed prior to a post-germination phase to obtain a protein product, wherein a yield of said protein product is at least about 1 g protein/kg seed.   
     
     
         2 . The method of  claim 1 , wherein the yield of said protein product is at least about 3 g protein/kg seed. 
     
     
         3 . The method of  claim 1 , wherein said plant seed belongs to a Fabaceae family. 
     
     
         4 . The method of claim  4 , wherein said plant seed that belongs to said Fabaceae family is a dry bean. 
     
     
         5 . The method of  claim 1 , wherein said plant seed belongs to a Poaceae family. 
     
     
         6 . The method of  claim 1 , wherein said plant seed belongs to said Brassicaceae family. 
     
     
         7 . The method of  claim 1 , wherein, in (a), (b), or (c), said plant seed is germinating. 
     
     
         8 . The method of  claim 1 , wherein said plant seed is non-germinating. 
     
     
         9 . The method of  claim 1 , wherein (b) comprises applying a bacteria to said plant seed comprising said protein expression vector. 
     
     
         10 . The method of claim  10 , wherein said bacteria comprises agrobacteria. 
     
     
         11 . The method of  claim 1 , wherein (b) comprises microbombardment. 
     
     
         12 . The method of  claim 1 , wherein said protein expression vector comprises an intron. 
     
     
         13 . The method of  claim 1 , wherein said protein comprises an antibody. 
     
     
         14 . The method of  claim 1 , wherein said protein comprises an enzyme. 
     
     
         15 . The method of  claim 1 , wherein said protein comprises a dairy protein. 
     
     
         16 . The method of  claim 15 , wherein said dairy protein is a casein protein. 
     
     
         17 . The method of  claim 16 , wherein said casein protein comprises αs1-casein, αs2-casein, β-casein, κ-casein, or a combination thereof. 
     
     
         18 . The method of  claim 1 , wherein said protein comprises an amino acid sequence with at least 80% sequence identity to SEQ ID NO:11. 
     
     
         19 . The method of  claim 1 , wherein said protein comprises an amino acid sequence with at least 80% sequence identity to SEQ ID NO:13. 
     
     
         20 . The method of  claim 1 , wherein said protein comprises an amino acid sequence with at least 80% sequence identity to SEQ ID NO:15. 
     
     
         21 . The method of  claim 1 , wherein said protein comprises insulin. 
     
     
         22 . The method of  claim 1 , wherein said protein comprises a signaling peptide. 
     
     
         23 . The method of  claim 1 , further comprising, in (d), extracting said protein from other proteins in said plant seed. 
     
     
         24 . The method of  claim 23 , wherein extracting said protein comprises his-tag purification. 
     
     
         25 . The method of  claim 23 , wherein extracting said protein comprises ion-exchange chromatography. 
     
     
         26 . The method of  claim 23 , wherein extracting said protein comprises using alkaline conditions. 
     
     
         27 . The method of  claim 23 , wherein extracting said protein comprises using acidic conditions. 
     
     
         28 . The method of  claim 1 , further comprising storing said plant seed at a temperature of between 0° C. and −40° C. after (c) and before (d).